A coupled optical enzyme assay for phosphopentomutase |
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| Authors: | Maria Grazia Tozzi Roberta Catalani Pier Luigi Ipata Umberto Mura |
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| Affiliation: | Institute of Biochemistry, Biophysics and Genetics, Faculty of Science, University of Pisa, Via A. Volta 4, 56100 Pisa, Italy |
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| Abstract: | Published assays for phosphopentomutase activity are based on acid lability differences between ribose 1-phosphate and ribose 5-phosphate. The present work describes a new method in which the isomerization of ribose 5-phosphate to ribose 1-phosphate is followed spectrophotometrically at 265 nm by coupling it with the following two-stage enzymatic conversion: ribose 1-phosphate + adenine ? phosphate + adenosine (adenosine phosphorylase); adenosine + H2O → inosine + NH3 (adenosine deaminase). The method has been used to show some properties of Escherichia coli phosphopentomutase. |
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